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Titolo:
Rat marrow stromal cells rapidly transduced with a self-inactivating retrovirus synthesize L-DOPA in vitro
Autore:
Schwarz, EJ; Reger, RL; Alexander, GM; Class, R; Azizi, SA; Prockop, DJ;
Indirizzi:
Tulane Univ, Hlth Sci Ctr, Ctr Gene Therapy, New Orleans, LA 70112 USA Tulane Univ New Orleans LA USA 70112 e Therapy, New Orleans, LA 70112 USA Med Coll Penn & Hahnemann Univ, Dept Neurol, Philadelphia, PA 19102 USA Med Coll Penn & Hahnemann Univ Philadelphia PA USA 19102 ia, PA 19102 USA Med Coll Penn & Hahnemann Univ, Dept Radiat Oncol, Philadelphia, PA 19102 USA Med Coll Penn & Hahnemann Univ Philadelphia PA USA 19102 ia, PA 19102 USA
Titolo Testata:
GENE THERAPY
fascicolo: 16, volume: 8, anno: 2001,
pagine: 1214 - 1223
SICI:
0969-7128(200108)8:16<1214:RMSCRT>2.0.ZU;2-1
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEIN; MESENCHYMAL STEM-CELLS; MEDIATED GENE-TRANSFER; HUMAN BONE-MARROW; TYROSINE-HYDROXYLASE; PARKINSONS-DISEASE; NONHEMATOPOIETIC TISSUES; HEMATOPOIETIC-CELLS; EXPRESSION; VECTORS;
Keywords:
rat marrow stromal cells; L-DOPA; GFP; expansion; retrovirus;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
34
Recensione:
Indirizzi per estratti:
Indirizzo: Prockop, DJ Tulane Univ, Hlth Sci Ctr, Ctr Gene Therapy, SL99,1430 Tulane Ave, New Orleans, LA 70112 USA Tulane Univ SL99,1430 Tulane Ave New OrleansLA USA 70112 USA
Citazione:
E.J. Schwarz et al., "Rat marrow stromal cells rapidly transduced with a self-inactivating retrovirus synthesize L-DOPA in vitro", GENE THER, 8(16), 2001, pp. 1214-1223

Abstract

Autologous bone marrow stromal cells engineered to produce 3.4-dihydroxyphenylalanine(L-DOPA) can potentially be used as donor cells for neural transplantation in Parkinson's disease. Here. we examined the possibility of using several different promoters and either a self-inactivating retrovirus (pSIR) or standard retroviruses to introduce into marrow stromal cells (MSCs), the two genes necessary for the cells to synthesize L-DOPA. pSIR vectors were constructed using the mouse phosphoglycerate kinase-1 (PGK) promoter or the cytomegalovirus (CMV) promoter to drive expression of either a GFP reporter gene or a bicistronic sequence containing the genes for human tyrosine hydroxylase type I (TH) and rat GTP cyclohydrolase I (GC) separated by an internal ribosome entry site (IRES). rMSCs were successfully transduced with both standard retroviral vectors and pSIR containing the PGK promoter. Transduced rMSCs expressed GFP (90.4-94.4% of cells) or were able to synthesize and secrete L-DOPA (89.0-283 pmols/10(6) cells/h). After transduced rMSCs were plated at low density (3-6 cells/cm(2)), the cells expanded over 1000-fold in 3-4 weeks, and the rMSCs continued to either express GFP or produce L-DOPA. Furthermore, two high-expressing clones were isolated and expanded at low-density from rMSCs transduced with pSIR driven by the PGK promoter (97.0% GFP+ or 1096.0 pmols L-DOPA/10(6) cells/h).

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Documento generato il 26/01/20 alle ore 09:58:46